RT-PCR Detection of Equine Arteritis Virus from Various Samples of an Experimentally Infected Pregnant Mare

Abstract

Various clinical and tissue materials derived from an experimentally infected mare were examined for the equine arteritis virus (EAV) by the RT-PCR method. For the reverse transcriptional step, a specific primer (R1 primer) and random (nonspecific) 9-mers primer were used. The specific 292 bp band could be detected clearly with the random primer, but not the specific primer from clinical and various autopsy samples of the pregnant mare and aborted fetus. The sensitivity to detect 7 strains of EAV by RT-PCR with the random primer was tested. The detection limit of Bibuna and Vienna strains of EAV was about 100 PFU, although that of other strains, Bucyrus, modified Bucyrus, 84KY-A1, Red Mile and Wroclaw ranged from 1 to 10 PFU. By our RT-PCR method, EAV could be detected within 1 day and could be obtained directly from a urine sample, from which the virus was difficult to isolate without pretreatment due to toxicity in cell culture.